Fundamental Guide to Liquid Chromatography Mass ...

1 Fundamental Guide toLiquid ChromatographyMass spectrometry (LCMS)C10G-E065 AMILESTONESFOR Liquid CHROMATOGRAPHYAND mass SPECTROMETRYS himadzu Corporation focus on analytical and measuring instruments, medical systems, aircraft equipment and industrial machinery. Together with our partners around the world, we diligently strive to be a company that meets the challenges faced by society and develop products which contribute to the growth of industries and the enrichment of peoples Shimadzu Sr. founds Shimadzu Corporation1987 WORLD S 1ST MALDI-MS LCMS-2010 AXIMA-CFRLAMS-50 KLCMS-2010 AAXIMA-QIT200220032018 NOBEL PRIZE IN CHEMISTRY AWARDED TO KOICHI TANAKAK oichi Tanaka mass spectrometry Research Laboratory established20042008201020142013200620152 016 World s 1st Ion trap TOF LCMSLCMS-IT-TOFW orld-Class General PurposeMALDI-7090 World s 1st Automated LCMS Module for Clinical ApplicationsUnified Chromatography (UC)

2 UHPLC/SFC Switching SystemCLAM-2000 MALDI-8020 Nexera MikrosDPiMS-2020 LAUNCH OF UFMS TECHNOLOGYLAUNCH OF UFMS TECHNOLOGYM inimized cross talk with UFsweeper Prominence UFLCXRO pening up new worlds and expanding possibilities through ultra-fast Liquid chromatographyAcquire mass spectra directly from crude samplesDelivers superior performance across a wider application range with flexibility and reliabilityWorld's 1st fully automated system that combines on-line SFE and SFC in a single flow path. Ultimate performance in identification and structural characterization of biomoleculesProvides high resolution morphological imaging overlaid with molecular distribution images Integrated LC systems that provide high speed analysis, simplified method transfer, minimized environmental impact and easy maintenanceQuick and easy MS analysis without sample preparationWith the added UF-Link, this low flow LCMS system improves operability and enables high-sensitivity Higher sensitivity with UF-Lens Improved CID efficiency with UFsweeperII Cutting-edge UFsweeperIII , UF-grating and UF-FlightTube allows high-speed data acquisition for high-throughput analysisAchieved UFswitching.

3 UFscanning and UF-MRM Next-levelquantitative detection with UF-Qarray Quadruopole Time-of-Flight Liquid Chromatography mass SpectrometerShimadzu s revolutionary technology bringing next-generation analysis performanceLCMS-2020 LCMS-8030 LCMS-8040 LCMS-8050 LCMS-8060 LCMS-8045 LCMS-9030 MALD-TOF MS modelsFor Research Use Only. Not for use in diagnostic publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of names, products/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation, its subsidiaries or its affiliates, whether or not they are used with trademark symbol TM or.

4 Third-party trademarks and trade names may be used in this publication to refer to either the entities or their products/services, whether or not they are used with trademark symbol TM or .Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own. The information contained herein is provided to you as is without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice. Shimadzu Corporation First Edition: October, 2018 Updated on: March, 2019 MILESTONES FOR Liquid Chromatography AND mass SPECTROMETRYMILESTON FRQURQSDLRC HRANG RC SCSPYDLASP SCT E1 SGNQLC9 LCGDQNE1 CDG8 E1 TLASP GYGD1EG8 SLQAQSHD 17 NLUE1CD SCT LCTNGDQLSP ESAILC1QY2 0R91DI1Q 3 LDI RNQ USQDC1QG SQRNCT DI1 3 RQPT8 31 TLPL91 CDPY GDQLK1 DR o1 S AREUSCY DISD E11DG DI1 AISPP1C91G HSA1T oY GRAL1DY SCT T1K1 PRU UQRTNADG 3 ILAI ARCDQLoND1 DR DI1 9QR3DI RH LCTNGDQL1G SCT DI1 1 CQLAIE1CD RH U1 RUP1Gi PLK1G21875 Genzo Shimadzu Sr.

5 Founds Shimadzu Corporation1987 WORLD S 1ST MALDI-MS LCMS-2010 AXIMA-CFRLAMS-50 KLCMS-2010 AAXIMA-QIT200220032018 NOBEL PRIZE IN CHEMISTRY AWARDED TO KOICHI TANAKAK oichi Tanaka mass spectrometry Research Laboratory established20042008201020142013200620152 016 World s 1st Ion trap TOF LCMSLCMS-IT-TOFW orld-Class General PurposeMALDI-7090 World s 1st Automated LCMS Module for Clinical ApplicationsUnified Chromatography (UC) UHPLC/SFC Switching SystemCLAM-2000 MALDI-8020 Nexera MikrosDPiMS-2020 LAUNCH OF UFMS TECHNOLOGYLAUNCH OF UFMS TECHNOLOGYM inimized cross talk with UFsweeper Prominence UFLCXRO pening up new worlds and expanding possibilities through ultra-fast Liquid chromatographyAcquire mass spectra directly from crude samplesDelivers superior performance across a wider application range with flexibility and reliabilityWorld's 1st fully automated system that combines on-line SFE and SFC in a single flow path.

6 Ultimate performance in identification and structural characterization of biomoleculesProvides high resolution morphological imaging overlaid with molecular distribution images Integrated LC systems that provide high speed analysis, simplified method transfer, minimized environmental impact and easy maintenanceQuick and easy MS analysis without sample preparationWith the added UF-Link, this low flow LCMS system improves operability and enables high-sensitivity Higher sensitivity with UF-Lens Improved CID efficiency with UFsweeperII Cutting-edge UFsweeperIII , UF-grating and UF-FlightTube allows high-speed data acquisition for high-throughput analysisAchieved UFswitching , UFscanning and UF-MRM Next-levelquantitative detection with UF-Qarray Quadruopole Time-of-Flight Liquid Chromatography mass SpectrometerShimadzu s revolutionary technology bringing next-generation analysis performanceLCMS-2020 LCMS-8030 LCMS-8040 LCMS-8050 LCMS-8060 LCMS-8045 LCMS-9030 MALD-TOF MS modelsForewordHirano IchiroLCMS Product ManagerWe are very happy to deliver you our first LCMS Primer which has very well compiled the basic principles and theory of mass spectrometry .

7 It also describes the history of development of various innovative technologies to enhance the performance of our LC-MS/MS systems. It serves both as a textbook and a practical application Guide , regardless of the scientific background and working field of the readers. All market figures indicate that Shimadzu is the fastest-growing mass spectrometry company in the last decade. Since the product launch of our first triple quadrupole LC-MS/MS, LCMS-8030 in September 2010, we have been continuously rolling out the new LC-MS/MS systems to keep up to date with increasing needs. This year (2018), we are celebrating our landmarking launch of the LCMS-9030, our first ever Q-TOF instrument. Given these achievements, it is now our obligation and responsibility to deliver a primer to increase general understanding of the scientific foundation that we stand upon, while also pointing to the future of the LCMS that we are to realize.

8 I would like to express my gratitude for your eagerness to learn more about LCMS and taking time to read this primer. I sincerely hope that it satisfies your enthusiasm and gets you prepared for the new era of 1 Introduction to LCMS 4 Fundamentals of LC, MS and LCMS 5 Comparison of LCMS and other techniques 9 Applications of LCMS 11 Chapter 2 Integrating LC and MS 12 Basic instrumentation of LCMS 13 Interfaces for LCMS 15 Electrospray Ionization (ESI) 16 Atmospheric Pressure Chemical Ionization (APCI) 18 Atmospheric Pressure Photoionization (APPI) 18 Overview of Atmospheric Pressure Ionization (API) 21 Mobile phases compatible for LCMS 24 Chapter 3 Principles of MS 26 Introduction to mass analyzers 27 Magnetic sector MS 28 Quadrupole MS 30 Time-of-Flight (TOF) MS 33 Ion Trap (IT)

9 MS 37 Comparison of mass analyzers 39 Chapter 4 Introduction to tandem MS systems 41 Core principles 42 Types of MS/MS systems and their key characteristics 46 Triple Quadrupole (TQ) MS 47 Quadrupole Time-of-Flight (Q-TOF) MS 47 Multistage MS (MSn)50 Ion Trap Time-of-Flight (IT-TOF) MS 50 Comparison of MS/MS systems 51 Chapter 5 Challenges and development of LCMS 53 Analytical challenges and current limitations of LCMS 54 Key developments in Shimadzu s LCMS 56 Ultra-Fast mass spectrometry (UFMS) 56 Clinical Laboratory Automation Module (CLAM) 58 Future trends of LCMS 59 Shimadzu s Comprehensive LC and MS Solutions 60 List of ContentsAbbreviationsAPCI Atmospheric Pressure Chemical IonizationAPI Atmospheric Pressure Ionization APPI Atmospheric Pressure PhotoionizationCID Collision-induced Dissociation CLAM Clinical Laboratory Automation ModuleDUIS Dual Ion Source EI Electron Ionization ESI Electrospray IonizationGC Gas ChromatographyGCMS Gas Chromatography mass SpectrometryHILIC Hydrophilic Interaction Liquid ChromatographyHPLC High-Performance Liquid ChromatographyICP-MS

10 Inductively Coupled Plasma mass SpectrometryIEC Ion Exchange ChromatographyIT Ion TrapIT-TOF Ion Trap Time-of-Flight LC Liquid ChromatographyLCMS Liquid Chromatography mass SpectrometryMALDI Matrix-Assisted Laser Desorption/IonizationMALDI-TOF Matrix-Assisted Laser Desorption/Ionization Time-of-Flight MRM Multiple Reaction MonitoringMS mass SpectrometryMS/MS tandem mass SpectrometryMW Molecular WeightNPLC Normal Phase Liquid ChromatographyPDA Photodiode Array DetectorQ-IT Quadrupole Ion TrapQ-TOF Quadrupole Time-of-Flight RPLC Reversed Phase Liquid ChromatographyRT Retention TimeSEC Size Exclusion ChromatographySIM Selected Ion MonitoringSRM Selected Reaction MonitoringTIC Total Ion CurrentTOF Time-of-FlightTQ Triple QuadrupoleUFMS Ultra-Fast mass SpectrometryUHPLC Ultra-High-Performance Liquid ChromatographyUV-VIS Ultraviolet-Visible Spectroscopy Symbols Microamu Atomic mass unitB Magnetic flux densityE Electric fielde Elementary chargeg Gram k Kilom mass m/z mass -to-charge ration Nano Pa PascalV